Clinical Trials
Shrivastava, J Clinic Trials 2011, 1:1
http://dx.doi.org/10.4172/jctr.1000102
Research Article
Open Access
A Pilot Clinical Trial to Evaluate the Efficacy of a Topical Antiviral
Osmotically Active Hypertonic Solution for the Treatment of Influenza
Virus Induced Sore Throat
Ravi Shrivastava*
VITROBIO Research Institute, ZAC de Lavaur 63500 Issoire, France
Abstract
Objectives: The presence of free virus particles and bacteria on the surface of the throat is the main cause of
initiating and maintaining viral throat infection. In the absence of any topical antiviral treatment, the objectives of this
study were to evaluate the efficacy and safety of some specific virus glycoprotein neutralizing and proteolytic enzyme
binding plant tannins in an osmotically active hypertonic solution (VB-Th4) for the treatment of sore throat.
Methods: 60-patients having acute sore throat were treated with VB-Th4 spray for a maximum period of
14-consecutive days. 43 patients in the Standard treatment (ST) group received other commonly used treatments.
Effect on clinical signs, duration of recovery and requirement for the use of antibiotics was evaluated.
Results: VB-Th4 significantly reduced the signs of sore throat, bacterial load as well as throat pain, irritation and
inflammation immediately after the first treatment with complete recovery in 90% patients within 7-days. The time
required for complete recovery and the use of antibiotics was significantly reduced in the VB-Th4 compared to the
ST group.
Conclusion: In the absence of any topical antiviral drug, topical virus glycoprotein and protease inhibitors open
a new therapeutic approach to treat throat viral infections.
Keywords: Influenza virus; Sore throat; Proteases; Glycoproteins;
Procyanidins
Introduction
Throat viral infections, particularly the influenza and parainfluenza
virus inducing upper respiratory tract infections, are very common in
human beings [1].
Initially only a few virus particles come in contact with the throat
mucus membrane, enter into the cells, multiply and millions of new
virus particles are then liberated topically on the throat surface. These
free virus particles now attack new healthy cells and weakens local
immunity which is followed by secondary bacterial infection leading to
pain, irritation, inflammation, fever and other signs of flu.
Influenza is an enveloped virus with a complex viral coat containing
several glycoproteins (Gps). Virus envelop Gps are transmembrane
proteins, anchored to the envelope by a hydrophobic domain. Virus
uses these glycoproteins to attach to the cell membrane and to enter
into the host cell [2]. The differences in the surface Gp structures confer
different morphology and antigenicity in the same family of virus [3].
Continuous discovery of new virus surface Gps, their role in host cell
infection and frequent viral mutations makes the development of
a specific topical antiviral drug nearly impossible. In addition to the
virus surface glycoproteins, topically liberated proteases on the throat
surface also play an important role in facilitating influenza virus entry
into the cell [4,5].
In the absence of any specific antiviral drug, vaccines are currently
considered the best antiviral therapy but requires continuous adaptation
due to regular changes in virus surface GPs. This is the reason why
almost all the currently available antiviral drugs are directed to interfere
with intracellular virus multiplication process but have no effect at all
on free virus particles present on the throat surface, which is the main
cause of throat infections.
Taking into consideration the protein nature of the proteases and
the virus Gps on one hand and the protein binding properties of certain
J Clinic Trials
ISSN: JCTR, an open access journal
plant tannins, the aim of our research was to identify all the Gps and
the proteases involved in the influenza virus infection and to search
corresponding tannins or the procyanidin (PCD) fraction of the tannin
having a strong capacity to bind with these proteins so as to stop topical
viral growth.
Tannins are abundant in the plant kingdom and are known to bind
with a wide range of protein molecules [6]. The procyanidin (PCD)
fraction of the tannins contains big phenolic compounds with multiple
structure units and may have selective protein binding properties
[7]. This multiple structures of tannins confer them the possibilities
to form strong permanent hydrogen bonds with a group of proteins.
We analyzed the protein binding capacity of above 300 different PCDs
by incubating PCDs with virus suspension or proteases followed by
evaluating virus titre in influenza virus sensitive MDCK (Madin –Darby
Bovine Kidney) cells as described by Shrivastava [8]. The selected plants
PCDs were then added in a hypertonic osmotically active solution [9]
as a cleaning agent on the throat surface to evacuate the bounded
protein-PCD complexes from the throat surface. After initial selection
of the best antiviral composition, a pilot clinical trial was conducted to
verify clinical efficacy of the preparation, coded as VB-Th4 in patients
suffering from acute viral sore throat. A parallel group of patients
receiving other commonly used treatments was incorporated in the
*Corresponding author: Dr. Ravi shrivastava, PhD, VITROBIO Research
Institute, ZAC de Lavaur 63500 Issoire, France, Tel +33 4 73 55 05 05, Fax +33 4
73 55 00 11; E-mail: [email protected]
Received November 08, 2011; Accepted December 14, 2011; Published
December 19, 2011
Citation: Shrivastava R (2011) A Pilot Clinical Trial to Evaluate the Efficacy of
a Topical Antiviral Osmotically Active Hypertonic Solution for the Treatment of
Influenza Virus Induced Sore Throat. J Clinic Trials 1:102. doi:10.4172/jctr.1000102
Copyright: © 2011 Shrivastava R. This is an open-access article distributed under
the terms of the Creative Commons Attribution License, which permits unrestricted
use, distribution, and reproduction in any medium, provided the original author and
source are credited.
Volume 1 • Issue 1 • 1000102
Citation: Shrivastava R (2011) A Pilot Clinical Trial to Evaluate the Efficacy of a Topical Antiviral Osmotically Active Hypertonic Solution for the
Treatment of Influenza Virus Induced Sore Throat. J Clinic Trials 1:102. doi:10.4172/jctr.1000102
Page 2 of 7
study as a Standard Treatment (ST) group to evaluate the duration of
complete recovery and the use of antibiotics, if any, in the two groups.
Materials and Methods
Test product preparation
More than 300 different PCDs from 128 tannin rich known plants
were prepared using the method described by B. Giner-Chavez et al
[10]. In short, initial tannin rich plant extract was obtained with an
aqueous organic solvent containing 70% acetone and 30% water.
The extracts were then successively passed through Sephadex LH-20
columns by progressively increasing the volume of methanol (60 x
88.5 cm) and the intended fractions were eluted to produce a dry solid.
The product was identified by mass spectrometry. The extracts mainly
contained procyanidin (epicatechin –catechin) B1, B2, B3 and C1
fractions between 60-80% depending upon the part of the plant used.
The proteases involved in enhancing influenza virus entry into the
cells, virus surface glycoproteins, and specific PCDs capable to bind
with one or more of these proteins were evaluated employing the
methods as described previously [8]. In short, to identify the proteases,
influenza virus sensitive MDCK cells were grown in 96-well tissue
culture plates (Corning, USA) in vitro in a protease free Dulbecco’s
Modified Eagle’s Medium (DMEM). Cells were infected with a
multiple Gp expressing human parainfluenza virus type 1 (ATCC-US,
Ref: VR 94, Strain C35) adapted for tissue culture, to calculate 50 and
100% tissue culture infective doses (TCID50 & 100). Proteases were then
added into the culture medium, either individually or in association, to
evaluate the effect on virus growth. Increase in virus growth indicated
participation of the protease(s) to enhance viral infection. To evaluate
virus protein binding properties of PCDs, a fixed concentration of
virus (TCID100) and/or proteases was pre-incubated with a specific
PCD, and the reduction in virus growth was evaluated compared to the
corresponding controls. Low virus growth indicated virus or protease
neutralization with the PCD. All active PCDs were then associated
in varying concentrations so as to obtain 100% inhibition of virus
infection in vitro. The concentration of each PCD required to inactivate
100% virus growth in a single well (surface area 0.328 cm2) of 96-well
culture plate was determined and the final association was designated
as VB-Th4-PCDs. The PCD association (0.78%) was incorporated in
an osmotically active hypertonic solution [11] containing glycerol
(74.27%), honey (12.0%), and water (12.95%) so as to apply between
4-6 mg PCDs per application or between 15-20 mg PCDs per day on
the throat surface. The hypothesis was that the selected antiviral PCD
association will neutralize virus GPs and inactivate the virus, other
protease specific PCDs will bind and inactivate virus entry enhancing
proteases, and the osmotically active hypertonic solution will help to
attract hypotonic liquid from the inner parts of the throat thereby
cleaning the throat surface from conjugated virus-PCD, protease-PCD
and other contaminants from the throat surface. This triple action
filmogen solution was filled in 30-ml aluminium containers caped with
a spray for throat application and labelled as VB-Th4.
Clinical trial
Location: An open, label, prospective, multicentric, pilot study was
conducted by the Nexus Clinical Research Pvt Ltd at Mumbai in India
between 07-2009 to 01-2011.
Ethical aspects: This pilot study was conducted only after
the approval of Institutional Review Board/ Independent Ethical
committee agreed by the Indian Council of Medical research (ICMR)
J Clinic Trials
ISSN: JCTR, an open access journal
respecting GCP (Good Clinical Practice) and following the principles
laid down in the declaration of Helsinki and amends thereafter. The
investigator institute is authorized to conduct clinical trials and is
regularly inspected by the regulatory authorities. Only the subjects who
gave informed consent were included in the study.
Inclusion and exclusion criteria: The main inclusion criteria
were 1. Participants having all clinical sign and symptoms of acute
and recent (less than 72h) sore throat pharyngitis such as exudate and
swelling on tonsils or cervical lymph nodes, fever above 100.4°F (38°c),
pain and absence of cough with probability of strep pharyngitis above
50% according to the Centor criteria [12]. Although, patients were not
assigned a specific ICD 10 code, most of the patients were close to the
symptoms specified under ICD 10-CM diagnosis code J02.9 and all
were positive for Streptococcus culture. 2. Male and female between
age groups of 18-65 years, 3. Normal blood biochemical profile for
liver and kidney parameters. 4. No history of adverse effect or allergies
to any ingredient used in the product composition, 5 Not under any
antibacterial or antiviral treatment before recruitment. 6. Given written
consent and willing to follow the protocol as recommended.
Randomization: Patients were at randomly assigned by the clinical
investigators in each research centre to obtain an approximate ratio of
2:3 for ST and active treatment group at arrival on physical basis.
Study design: Patients in the VB-Th4 group were asked to spray
the solution over the throat surface every 20-30 minutes during the 2-3
hours at the beginning of the treatment and 3-4 times per day thereafter
up to complete recovery or up to a maximum period of 14 consecutive
days. ST group patients followed treatment recommendations advised
by their medical advisor.
Efficacy and safety evaluation: After recruitment, the medical
history of patient was recorded in the observation file. Upon evaluation
of the entire baseline parameters as per study protocol, each patient
received the test product and symptom observation diary.
At each time point, all the observations were recorded by an
experienced ENT clinical research coordinator or the clinical
investigator in collaboration with the patient at each clinical research
center.
As the main objective of the study was to measure throat
symptom relief efficacy of the test product, the patients treated with
VB-Th4 were evaluated for the intensity of throat pain, local throat
irritation, and throat redness on a commonly used clinical parameter
scoring scale [13] of 0 to 10 where 0 signifies no symptoms, 1-3 mild
symptoms, 4-6 moderate symptoms, and 7-10 severe symptoms. Being
an osmotically active solution engendering outward flow of liquid
and due to continuous deglutition movements of throat, it was not
possible to quantify virus amount on the throat surface. Nevertheless,
throat swabs were collected to quantify bacterial load by counting the
number of colony forming units (cfu/cm2), before 1st treatment, 2h
after 1st treatment and then on days 4, 7, 10 and 14 or up to complete
recovery. Throat swabs were collected even if the patient was found
negative during the previous examination to confirm the results but
only positive smears were used to calculate mean values. Results were
compared with before treatment values. To verify eventual systemic
reactions due to product application, complete hematological and
blood biochemical analyses were also performed at the start and at the
end of the study.
ST group patients were only checked for the presence of bacterial
acute sore throat (throat swab before recruitment), were allowed to
Volume 1 • Issue 1 • 1000102
Citation: Shrivastava R (2011) A Pilot Clinical Trial to Evaluate the Efficacy of a Topical Antiviral Osmotically Active Hypertonic Solution for the
Treatment of Influenza Virus Induced Sore Throat. J Clinic Trials 1:102. doi:10.4172/jctr.1000102
Page 3 of 7
Results
take any treatment prescribed by their clinical ENT specialist and were
asked to note any use of systemic antibiotics, duration of antibiotic
treatment if any, and the day of complete recovery, if applicable, during
the study period. All the participants of both groups were authorized to
take systemic antibiotics, if found necessary by the investigator.
Population analyses
Being a pilot clinical trial, a total 134 patients suffering from
acute sore throat were initially screened for the study in four different
hospitals in India. As shown in the flow diagram (Figure 4), 17 did not
meet inclusion criteria, 6 refused to participate and 2 were unable to
come for check up. Among 109 patients enrolled, 63 were at randomly
assigned to the VB-Th4 group among which 2 were lost during follow
up and 1 did not returned complete data. Among 60 patients who
followed complete VB-Th4 protocol, 36 (60.0%) were men and 24
(40.0%) women, between age group of 19-63 years (mean age 46.01
± 12.84 years), and had a mean body weight of 67.31 kg (± 12.82 kg).
Patients from both groups were considered completely recovered
when throat surface bacterial count was normal and all the key
symptoms of throat infection (pain, irritation, and redness) were
absent. Presence of minor coughing was not considered as a key
recovery parameter as slight cough always persists even in recovered
subjects.
The data obtained from 60 subjects of the VB-Th4 group who
completed the study were assessed for the safety and efficacy parameters
compared to the results obtained before the start of the study.
Among 46 other patients included in the ST group, 3 were lost
during follow up. Out of 43 results analyzed, 19 (44.18%) were men
and 24 (55.81%) were women. All subjects were positive for bacterial
cultures at the time of recruitment and the identified organism was
mainly Streptococcus. The 43 ST group subjects were taking either
one or more treatments such as the analgesics and antipyretics
(69.7%), topical antiseptics (48.8%), anti-inflammatory drugs (32.6%),
Ayurvedic plant drugs (20.93%) or gargling with warm salted water
(16.2%). This group served only to determine the duration of complete
recovery and the use of antibiotics.
Statistical methods used for data analysis
Clinical data were analyzed using SAS 9.1.3. Statistical program.
The statistical paired sample t-test, Wilcoxon Sign Rank tests were
used to compare the laboratory parameter. The Wilcoxon Sign Rank
test was used when the normality assumption was false. Descriptive
Statistics i.e. Mean Standard Deviation (SD), minimum and maximum
frequency distribution were used for the analysis of the demographic
details, clinical evaluation, medical history, and laboratory parameters.
If the P-value was greater than 0.05, the results were considered to be
not significant.
Presence or absence of Strep throat
On day-1 before treatment as well as 60 min after the 1st application
Before treatment
Day 1
Day 1
30-min after 1st application
Day 1
Presence of bacteria
60
60
60
Mean Bacterial count (cfu/cm²) ± SD
>1950
± 179.43
1187.20
± 127.28
745.60
±39.84
Day 4
20
(p=0.0098)
374.90
±15.50
Day 7
Day 10
Day 14
17
(p=0.0008)
N
N
N
N
N
N= Normal values
n= 60 patients
Table 1: Number of patients showing positive cultures for strep throat and mean bacterial count (cfu/cm² ± SD) in VB-Th4 group.
60
50
40
30
20
10
0
Before treatment
Day 1 - 2H after 1st
application
Day 4
No pain
Mild
Moderate
Day 7
Day 14
Severe
Figure 1: Number of patients evaluating the intensity of throat pain on a 0 to 10 scoring scale when 0 signifies no pain, 1-3 mild pain, 4-6 moderate pain 7-10 severe
pain before treatment, 2h after first product application and on the days 4, 7, 10 and 14 in the VB-Th4 group (n=60).
J Clinic Trials
ISSN: JCTR, an open access journal
Volume 1 • Issue 1 • 1000102
Citation: Shrivastava R (2011) A Pilot Clinical Trial to Evaluate the Efficacy of a Topical Antiviral Osmotically Active Hypertonic Solution for the
Treatment of Influenza Virus Induced Sore Throat. J Clinic Trials 1:102. doi:10.4172/jctr.1000102
Page 4 of 7
of VB-Th4, all the patients were positive for the presence of bacterial
throat infection, the predominant agent was Streptococcus pyogenes
in all the cases. Only 20/60 patients on the day 4 and 17/60 on the day
7 showed presence of bacteria above normal limits. All patients had a
normal bacterial count from the day 10 onwards. (Table 1)
Number of bacteria on the throat surface
The number of bacteria measured before VB-Th4 application
in throat swabs was very high and above the counting limits of 1950
(±179.43) cfu/cm2. 2h after after 1st product application, the mean
bacterial count was reduced to 1887.2 (±127.28) cfu/cm2 showing a
very rapid onset in bacterial reduction. The values were 745.6 (± 39.84)
cfu/cm2 on the day 4 and 374 (± 39.84 ) cfu/cm2 on day 7 with normal
values ( 50-100 cfu/cm2) from the day 10 onwards.(Table 1) This instant
and marked reduction in bacterial presence on the throat surface may
have been related to the hypertonic osmotically active properties of the
basic solution attracting hypotonic liquid from the inner surface of the
throat mucosa thereby detaching the microorganisms present on the
throat surface.
Effect on pharyngitis symptoms
Almost all the participant in both groups had moderate to severe
throat pain, throat irritation and inflammation at the start of treatment.
These parameters were evaluated only in the VB-Th4 group.
Effect on throat pain
57/60 patients had moderate to severe throat inflammation at the
start of the study (Figure 1). A marked analgesic effect was observed,
within 2h after the 1st test product application as only 2 patient recorded
severe pain and the pain intensity was reduced to moderate (34/60) or
mild (23/60) in other patients. The number of patients with moderate
pain was slightly increased on the day 4 (40 instead of 34 noticed 2h
after the 1st application) but was constant in the severe pain group. A
significant change was seen from the day 7 where 91% patients (55/60)
had only mild pain and on the day 10 where most of the patients
(58/60) had no throat pain.
These results show a very strong initial analgesic type of effect of
the test product followed by progressive reduction between days 1 and
4. As most of the patients (40/60) had nearly no bacterial infection just
after the 1st product application, it can be concluded that although VBTh4 exert a nearly instant anti-bacterial effect, the effects of VB-Th4
on the throat pain reduction are not instant. This may be related to the
fact that pain is usually a consequence of topical inflammation which
diminishes progressively.
Effect on throat irritation
As shown in the Figure 2, on the day 1 before treatment, 32/60
patients had severe and 24/60 patients had moderate symptoms of
throat irritation. This sensation was reduced in most of the patients
just after the 1st application of the product but was not completely
disappeared. A progressive reduction in the throat irritation score was
observed up to the day 7 when almost all the patients (55/60) were
completely recovered.
Effect on throat redness/inflammation
The throat inflammation was progressively diminished during the
first 4 days of treatment and only 6/60 patients had severe to moderate
inflammation from the day 7 onwards. (Figure 3) The results of this
parameter correspond to the concomitant reduction of pain and
bacterial infection observed in the same patients.
Complete recovery period compared to the ST group
As shown in the Table 2, the number of patients who stopped all
treatments after 2 days as they felt completely recovered was 31% in the
VB-Th4 group (n=60) compared to only 11% in the ST group (n=43).
On the day 7 of treatment, 61% participants in the VB-Th-4 group
stopped treatment compared to 25% in the ST group. On the day 10
almost all the patients (95.0%) in the VB-Th4 group stopped treatment
(57/60) compared to 28/43 patients (65.1%) in the ST group. Only 1
patient was not completely recovered in the VB-Th4 group on the day
14 and still had moderate to severe symptoms of throat pain, irritation,
and inflammation without the presence of bacterial infection. This
patient was diagnosed for pneumonia and was kept on antibiotherapy
thereafter. 9/43 patients in the ST group were not completely recovered
60
50
40
30
20
10
0
Before treatment
Day 1 - 2H after 1st
application
Day 4
None
Day 7
Mild
Moderate
Day 10
Day 14
Severe
Figure 2: Number of patients scoring the intensity of throat irritation on a 0 to 10 scoring scale (0 = no irritation, 1-3 = mild irritation, 4-6 = moderate irritation, 7-10 =
severe irritation) just before treatment, 2h after first product application and on the days 4, 7, 10 and 14 in the VB-Th4 group (n=60).
J Clinic Trials
ISSN: JCTR, an open access journal
Volume 1 • Issue 1 • 1000102
Citation: Shrivastava R (2011) A Pilot Clinical Trial to Evaluate the Efficacy of a Topical Antiviral Osmotically Active Hypertonic Solution for the
Treatment of Influenza Virus Induced Sore Throat. J Clinic Trials 1:102. doi:10.4172/jctr.1000102
Page 5 of 7
on the day 14 indicating that the recovery is much faster in the VBTh4 group especially when the treatment was started during the early
phase of throat infection. These results correspond to the absence of
bacterial infection observed in most of the patients right after the 2nd
day of treatment.
Antibiotherapy in the VB-Th4 compared to the ST group
during the study
As indicated in the Table 3, the number of patients requiring
antibiotic therapy was much higher in the ST group compared to the
VB-Th4 group. During the 14-day study period, only 4/60 patients
60
50
40
30
20
10
0
Before treatment
Day 1 - 2H after 1st
application
Day 4
Day 7
Day 10
Day 14
Figure 3: Intensity of throat redness/inflammation evaluated by the None
investigator
a 0 to 10
scale (0 = normal colour, 1-3 = slight, 4-6 = moderate, 7-10 = severe
Mild on
Moderate
Severe
inflammation) during the study period in the VB-Th4 group (n=60).
FLOW DIAGRAM:
Enrollment
Assessed for eligibility (n=134)
Excluded (n=25)
 Not meeting inclusion
criteria (n=17)
 Declined to participate (n=6)
Other reasons (n=2)
Randomized (n=109)
VB-Th4 group (n=63)
Allocation
Follow-Up
Placebo group (n=46)
Received allocated intervention (n= )
Did not receive allocated intervention
(give reasons) (n= )
Lost to follow-up (n=2)
Lost to follow-up (n=3)
Discontinued intervention (n=1)
Discontinued intervention (n=0)
Analysis
Analysed (n=60)
 Excluded from analysis (n=0)
Analysed (n=43)
 Excluded from analysis (n=0)
Figure 4:
J Clinic Trials
ISSN: JCTR, an open access journal
Volume 1 • Issue 1 • 1000102
Citation: Shrivastava R (2011) A Pilot Clinical Trial to Evaluate the Efficacy of a Topical Antiviral Osmotically Active Hypertonic Solution for the
Treatment of Influenza Virus Induced Sore Throat. J Clinic Trials 1:102. doi:10.4172/jctr.1000102
Page 6 of 7
(6.66%) in the VB-Th4 group required antibiotic therapy for an
average duration of 7.1 day compared to 14/43 patients (32.56%) in
the ST group for an average period of 9.8 days. On the day 14, only
1/60 patients in the VB-Th4 group was still continuing antibiotherapy
compared to 7/43 (16.23%) in the ST group. These results show that the
probable rate of serious complications requiring antibiotic therapy is
markedly reduced in the VB-Th4 group with respect to the number of
patients and the duration of treatment.
Other observations
No side effects or any undesirable reaction was observed in any of
the patients. None of the haematological, blood biochemical, or renal
parameters was affected in the VB-Th4 group. Most of the patients felt
an increase of liquid secretion in the mouth and topical heating effect
just after the application of the test product, which lasted for a short
duration (about 5-10 minutes).
Discussion
Viral and bacterial throat infections are usually seasonal epidemics
all over the world affecting 5-15% of the population [14]. This imposes
a considerable economic burden in the form of hospital and other
health care costs and lost productivity. In most of the cases, initial
viral infection, particularly the influenza virus, is the main cause. After
initial virus contact with the upper respiratory tract mucus membrane,
virus multiplies in a few cells and cell lyses liberates a large amount
of free virus particles on the throat surface. These newly liberated
virus particles attach new healthy cells, multiplies, and weakens local
immunity followed by secondary bacterial infection. Viral throat
infection usually leads to sore throat which is characterized by throat
pain, discomfort, or throat irritation, running nose, and cough. These
symptoms may be followed or associated with Strep throat, also known
as Streptococcal pharyngitis or Streptococcal sore throat, caused by
group A Streptococcus bacteria resulting in fever, patches of pus on the
throat surface, and swollen lymph nodes [1]. Patients opt for treatment
only during later stage when bacterial infection and clinical signs of
pharyngitis are well established. Symptoms may last for 2-3 weeks in
untreated patients.
In the absence of any effective topical antiviral drug, intracellular
antivirals can be used but they take at least 3-4 days to exert their effect
Group
Before treatment
N° VB-Th4
% population
N° standard treatment (ST) group
% population
and are of no use except to reduce the duration of symptoms by one or
two days [15]. Although these drugs are very effective to prevent the
viral infection, they are expensive, have many side effects, and patients
are usually reluctant to take any preventive medication. Antiviral
vaccines are useful but due to frequent minor genetic changes, known
as antigenic drift, vaccines require annual reformulation and can be
used only in the developed countries [16,17]. Therefore treatments
such as topical antiseptics, expectorants, local anesthetics, mucolytic
drugs, or salt water gargles are commonly used to relieve symptoms.
Antibiotic therapy is employed in nearly 50% sore throat patients
for an average duration of 10 days to prevent or to cure secondary
Streptococcus infection [18] but all these treatments have no effect
on millions of free virus particles present on the throat surface and
responsible for initiating and maintaining the infection.
Therefore, the scientific approach to treat viral throat infection lies
in neutralizing the virus particles on the throat surface, in inhibiting
viral entry into the host cells, and in removing bacterial infection
simultaneously.
Unfortunately, due to the complexity of the influenza virus
envelop with at least 16 haemagglutenin and 9 neuraminidase Gps,
due to simultaneous involvement of different proteases in the viral
entry [4] and subsequent secondary bacterial infection, it is practically
impossible to find a single drug with can act on all these parameters at
a time.
As the virus surface Gps and proteases are protein in nature and
as certain specific plant PCDs have a strong affinity to bind with the
proteins, the aim of our research was to employ the PCDs as topical
non specific antiviral agents. Virus Gps and virus entry enhancing
proteases were identified by incubating more than 350 individual PCDs
with virus or with proteases and analyzing their effect on virus growth
in vitro. Only 8 among 350 PCDs were found active and only 2/8 PCDs
isolated from Vitis vinifera seeds and Sambuscus nigra fruits were
capable to inhibit virus growth above 90% in vero cells. These PCDs
(0.78%) were incorporated in a hypertonic, viscous solution so as to
generate an osmotically active film over the throat surface attracting
hypotonic liquid from the throat mucosal surface and detaching the
bacteria due to this mechanical effect. Honey was added to improve the
viscosity and anti microbial activity of VB-Th4. Such a product could
Day 1
Day 2
Day 7
Day 10
Day 14
60
0
19
31.66
37
61.66
57
95.00
59
98.33
5
11
28
34
43
0
11.62
25.58
65.11
79.06
The patients not recovered completely after the day 14 were not followed. The difference is statistically significant compared to the ST group (p<0.05) from the day
2 onwards.
a:
Table 2 Number of patients who stopped treatment due to complete recovery during the study.a
Group
Before treatment
N° VB-Th4
% population
60
N° standard treatment (ST) group
% population
43
Day 1
Day 2
Day 7
Day 10
Day 14
0
2
3.33
3
5.0
4
6.66%
1
1.66
2
13
14
8
7
4.65%
30.23
32.56
18.60
16.28
The patients not recovered completely after the day 14 were not followed. The difference is statistically significant compared to the ST group (p<0.05) from the day 1
onwards.
a:
Table 3 Number of patients recommended antibiotherapy by their medical doctor from the date of entry into the study.a
J Clinic Trials
ISSN: JCTR, an open access journal
Volume 1 • Issue 1 • 1000102
Citation: Shrivastava R (2011) A Pilot Clinical Trial to Evaluate the Efficacy of a Topical Antiviral Osmotically Active Hypertonic Solution for the
Treatment of Influenza Virus Induced Sore Throat. J Clinic Trials 1:102. doi:10.4172/jctr.1000102
Page 7 of 7
have topical virus neutralizing, protease inhibitor and antibacterial
activities simultaneously.
The results of this study clearly show that VB-Th4 exert nearly
instant effects to reduce the number of bacteria on the throat surface.
The rapidity of anti-bacterial effect indicates a physical effect of outward
exudation of hypotonic liquid, creating a flux of liquid towards the
outer part of the throat surface and subsequent bacterial detachment.
Patients comments concerning heat sensation on the throat surface
and excessive liquid flow from the throat during the first 5-10 minutes
after each product application also confirms this mode of action.
Although the amount of virus particles on the throat surface was
difficult to measure, it is very likely that the free virus particles were
also cleaned from the throat surface reducing or stopping new throat
cell infection. The marked reduction observed in the symptomatic
parameters of throat infection such as pain, reddish color indicating
inflamed mucosa, and throat irritation also indicate absence of new
virus infection which may have been related to PCD binding with
the virus Gps and proteases. Experiments conducted during the
research phase indicated that the selected PCDs neutralize above 80%
of influenza virus particles and the proteases when PCDs were preincubated for 30-minutes with virus suspension or selected proteases
before vero cell infection in vitro [8]. Reduction in throat bacterial
count and new viral infection helps rapid recovery thereby reducing
the requirement of antibiotic therapy by nearly 50% compared to the
ST group.
No side effects or local reaction was noticed in any of the patients,
suggesting that VB-Th4 solution is a safe and nearly instant therapy
for the virus originated throat infections. Due to the mechanical mode
of action of this product, VB-Th4 can be considered a non specific
treatment for the throat infections of viral or bacterial origin. In the
absence of any topical, safe, inexpensive and effective topical antiviral
drug, the use of topical viral Gp inhibitors in association with protease
inactivators may constitute a new promising therapeutic approach for
viral and bacterial throat infections.
8. Shrivastava R (2011) A new therapeutic approach to neutralize throat surface
proteases and virus glycoproteins simultaneously for the treatment of influenza
virus infection. Int Journal Virology DOI: 10.3923.
9. Shrivastava R (2005) Non-solid composition for local application. PCT PatentWO 00/74668. Published 01.07.2002. US 6, 964,783 B1.
10.Giner-Chavez BI, Van Soest PJ, Robertson JB, Lascano C, Reed JD, et al.
(1997) A Method for Isolating Condensed Tannins from Crude Plant Extracts
with Trivalent Ytterbium. J Sci Food Agric 74: 359–368.
11.Shrivastava R (2011) New synergistic composition for the treatment of topical
viral infections. PCT Patent WO 2011/082835 A1, published 14.07.2011.
12.Farrar JT, Polomano RC, Berlin JA, Strom BL (2010) A comparison of change
in the 0-10 numeric rating scale to a pain relief scale and global medication
performance scale in a short-term clinical trial of breakthrough pain intensity.
Anesthesiology. 112: 1464-1472.
13.Wagner FP, Mathiason MA (2008) Using centor criteria to diagnose
streptococcal pharyngitis. Nurse Pract 33: 10-12.
14.Clark NM, Lynch JP (2011) Influenza: epidemiology, clinical features, therapy,
and prevention. Semin Respir Crit Care Med 32: 373-392.
15.Lin P, Torres G, Tyring SK (2003) Changing paradigms in dermatology:
antivirals in dermatology. Clin Dermatol 21: 426-446.
16.Carter NJ, Curran MP (2011) Live Attenuated Influenza Vaccine (FluMist®;
Fluenz™): A Review of its Use in the Prevention of Seasonal Influenza in
Children and Adults. Drugs 71: 1591-1622.
17.Petrie JG, Ohmit SE, Johnson E, Cross RT, Monto AS (2011) Efficacy studies
of influenza vaccines: effect of end points used and characteristics of vaccine
failures. J Infect Dis 203:1309-1315.
18.Altamimi S, Khalil A, Khalaiwi KA, Milner R, Pusic MV, et al. (2009) Short
versus standard duration antibiotic therapy for acute streptococcal pharyngitis
in children. Cochrane Database Syst Rev CD004872.
Acknowledgments
This study was sponsored by the Vitrobio Research Institute in France. We
sincerely thank Dr. Meghna Kotak Bhatt for her valuable guidance and suggestions
during this study and Ms. Nathalie Cucuat for her technical assistance. There is no
conflict of interest or sources of funding for the author.
References
1. West JV (2002) Acute upper airway infections. Br Med Bull 61: 215-230.
2. Kido H, Okumura Y, Takahashi E, Pan HY, Wang S, et al. (2012) Role of host
cellular proteases in the pathogenesis of influenza and influenza-induced
multiple organ failure. Biochim Biophys Acta 1824: 186-194.
3. Gravel C, Li C, Wang J, Hashem AM, Jaentschke B, et al. (2011) Quantitative
analyses of all influenza type A viral hemagglutinins and neuraminidases using
universal antibodies in simple slot blot assays. J Vis Exp doi: 10.3791/2784.
4. Kido H, Okumura Y, Takahashi E, Pan HY, Wang S, et al. (2008) Host envelope
glycoprotein processing proteases are indispensable for entry into human cells
by seasonal and highly pathogenic avian influenza viruses. J Mol Genet Med
3: 167-175.
5. Delboy MG, Roller DG, Nicola AV (2008) Cellular proteasome activity facilitates
herpes simplex virus entry at a postpenetration step. J Virol 82: 3381-3390.
6. Tarahovsky YS (2008) Plant polyphenols in cell-cell interaction and
communication. Plant Signal Behav 3: 609-611.
7. Zhang L, Chen J, Wang Y, Wu D, Xu M (2010) Phenolic extracts from Acacia
mangium bark and their antioxidant activities. Molecules 15: 3567-3577.
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